SUMMARY Staphylococcus aureus, a major human pathogen, has a collection of virulence factors and the ability to acquire resistance to most antibiotics. This ability is further augmented by constant emergence of new clones, making S. aureus a “superbug.” Clinical use of methicillin has led to the appearance of methicillin-resistant S. aureus (MRSA). The past few decades have witnessed the existence of new MRSA clones. Unlike traditional MRSA residing in hospitals, the new clones can invade community settings and infect people without predisposing risk factors. This evolution continues with the buildup of the MRSA reservoir in companion and food animals. This review focuses on imparting a better understanding of MRSA evolution and its molecular characterization and epidemiology. We first describe the origin of MRSA, with emphasis on the diverse nature of staphylococcal cassette chromosome mec (SCCmec). mecA and its new homologues (mecB, mecC, and mecD), SCCmec types (13 SCCmec types have been discovered to date), and their classification criteria are discussed. The review then describes various typing methods applied to study the molecular epidemiology and evolutionary nature of MRSA. Starting with the historical methods and continuing to the advanced whole-genome approaches, typing of collections of MRSA has shed light on the origin, spread, and evolutionary pathways of MRSA clones.

Methicillin-Resistant Staphylococcus aureus: Molecular Characterization, Evolution, and Epidemiology

Quaternary ammonium compounds (QACs) have represented one of the most visible and effective classes of disinfectants for nearly a century. With simple preparation, wide structural variety, and versatile incorporation into consumer products, there have been manifold developments and applications of these structures. Generally operating via disruption of one of the most fundamental structures in bacteria-the cell membrane-leading to cell lysis and bacterial death, the QACs were once thought to be impervious to resistance. Developments over the past decades, however, have shown this to be far from the truth. It is now known that a large family of bacterial genes (generally termed qac genes) encode efflux pumps capable of expelling many QAC structures from bacterial cells, leading to a decrease in susceptibility to QACs; methods of regulation of qac transcription are also understood. Importantly, qac genes can be horizontally transferred via plasmids to other bacteria and are often transmitted alongside other antibiotic-resistant genes; this dual threat represents a significant danger to human health. In this review, both QAC development and QAC resistance are documented, and possible strategies for addressing and overcoming QAC-resistant bacteria are discussed.

Quaternary Ammonium Compounds: An Antimicrobial Mainstay and Platform for Innovation to Address Bacterial Resistance

Summary In Asia, most reports on the epidemiology of community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA) are from developed countries, with few data from resource-limited countries, not because of low actual prevalence, but probably because of scarce diagnostic facilities. The rate of MRSA in all community-associated S aureus infections in Asian countries ranges from 2·5% to 39%. Unlike the predominance of USA300-sequence type (ST) 8 staphylococcal cassette chromosome mec (SCC mec ) type IV in the USA, the molecular epidemiology of CA-MRSA in Asia is characterised by clonal heterogeneity, similar to that in Europe. The emergence of CA-MRSA is a threat in both community and hospital settings because such strains are now more prevalent than are health-care-associated MRSA (HA-MRSA) strains. Many epidemic clones are in circulation in Asia and with scarce data available, concern has arisen that CA-MRSA could have devastating results if it becomes epidemic in resource-poor regions. The epidemiology of CA-MRSA in Asia is closely linked with the health of both developing and developed countries. The present situation of CA-MRSA in Asia is important not only for local public health, but also to provide a better understanding of the successful epidemic clones of this global pathogen.

https://www.thelancet.com/pdfs/journals/laninf/PIIS1473-3099(13)70136-1.pdf

Molecular epidemiology of community-associated meticillin-resistant Staphylococcus aureus in Asia

http://www.cleaning-for-health.org/files/2011/08/community-acquired-methicillin.pdf

Community-acquired methicillin-resistant Staphylococcus aureus: community transmission, pathogenesis, and drug resistance

A total of 108 S. aureus isolates from 16 major hospitals located in 14 different provinces in China were characterized for the profiles of 18 staphylococcal enterotoxin (SE) genes, 3 exfoliatin genes (eta, etb and etd), and the toxic shock syndrome toxin gene (tsst) by PCR. The genomic diversity of each isolate was also evaluated by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and accessory gene regulator (agr) typing. Of these strains, 90.7% (98/108) harbored toxin genes, in which tsst was the most prevalent toxin gene (48.1%), followed by sea (44.4%), sek (42.6%) and seq (40.7%). The see and etb genes were not found in any of the isolates tested. Because of high-frequency transfer of toxin gene-containing mobile genetic elements between S. aureus strains, a total of 47 different toxin gene combinations were detected, including a complete egc cluster in 19 isolates, co-occurrence of sea, sek and seq in 38 strains, and sec and sel together in 11 strains. Genetic typing by PFGE grouped all the strains into 25 clusters based on 80% similarity. MLST revealed 25 sequence types (ST) which were assigned into 16 clonal complexes (CCs) including 2 new singletons. Among these, 11 new and 6 known STs were first reported in the S. aureus strains from China. Overall, the genotyping results showed high genetic diversity of the strains regardless of their geographical distributions, and no strong correlation between genetic background and toxin genotypes of the strains. For genotyping S. aureus, PFGE appears to be more discriminatory than MLST. However, toxin gene typing combined with PFGE or MLST could increase the discriminatory power of genotyping S. aureus strains.

/pdf/genotypes-and-toxin-gene-profiles-of-staphylococcus-aureus-377zyieh75.pdf

Genotypes and Toxin Gene Profiles of Staphylococcus aureus Clinical Isolates from China

The molecular epidemiology and antimicrobial susceptibilities of 273 Staphylococcus aureus isolates positive for the exfoliative toxin-encoding gene obtained from patients with impetigo in Japan in 2006 were studied. The mecA gene was detected in 74 meticillin-resistant S. aureus (MRSA) and 23 meticillin-susceptible S. aureus (MSSA) isolates. All isolates with the staphylococcal cassette chromosome (SCC) mec were classified into type IV (92.8%, 90/97) or V (7.2%, 7/97). The ET-encoding gene etb was found primarily in strains with mecA (87.7%, 71/81), whilst eta (86.6%, 161/186) was detected mainly in strains without mecA. The chromosomal enterotoxin-encoding gene cluster egc was found in 83.0% of strains with eta, whilst no enterotoxin-encoding gene was detected in strains with only etb. PFGE showed that each strain carrying eta, etb and etd could be classified into distinct groups. The susceptibility profiles of MRSA to antimicrobial agents excluding beta-lactams were similar to those of MSSA. Gentamicin- and clarithromycin-resistant strains were frequently found for both MRSA and MSSA. The aminoglycoside-resistance gene aacA-aphD was detected in 97.3% of MRSA and 85.4% of MSSA. Additionally, the macrolide-resistance gene ermA or ermC was detected in 67.6% of MRSA and 71.4% of MSSA. Therefore, these results suggest that SCCmec types IV or V have spread, particularly in MSSA carrying etb in the community.

Molecular epidemiology and antimicrobial susceptibilities of 273 exfoliative toxin-encoding-gene-positive Staphylococcus aureus isolates from patients with impetigo in Japan.

Recently, the USA300 clone, which is a Panton-Valentine leukocidin (PVL)-positive clonal complex 8-staphylococcal cassette chromosome mec type IV (CC8-IV) community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) strain, emerged in community and hospital settings in Japan. Hence, clonal types of CA-MRSA strains are predicted to be changing. Nonetheless, long-term surveillance of CA-MRSA has not been conducted in Japan. Here, we investigated the transition and current status of CA-MRSA strains isolated from outpatients with impetigo; the samples were collected between 2007 and 2016 in Kagawa, Japan. The detection rate (22.8%, 488/2139 strains) of MRSA slightly decreased in these 10 years. Molecular epidemiological analyses showed that the prevalence of the CC89-II clone, which is a typical CA-MRSA genotype of causative agents of impetigo, significantly decreased from 48.0% (48/100 strains) in 2007-2009 to 21.9% (16/73 strains) in 2013-2016. By contrast, a non-USA300 CC8-IV clone, which is a highly pathogenic CA-MRSA/J clone, significantly increased in prevalence from 9.0% (9/100 strains) to 32.9% (24/73 strains). The prevalence of PVL-positive CA-MRSA strains increased annually from 2012 (0%) to 2015 (6.7%), whereas only one of these strains turned out to be the USA300 clone. Antibiotic susceptibility data revealed that the rates of resistance to gentamicin and clindamycin among CA-MRSA strains decreased along with the decreased prevalence of the CC89-II clone and increased prevalence of the CA-MRSA/J clone. Our data strongly suggest that the clonal types and antibiotic susceptibility of CA-MRSA isolated from patients with impetigo dramatically changed during the last 10 years in Japan.

Clonal change of methicillin-resistant Staphylococcus aureus isolated from patients with impetigo in Kagawa, Japan.

Cannabinoid Type 1 Receptors in the Basolateral Amygdala Regulate ACPA-Induced Place Preference and Anxiolytic-Like Behaviors

Cannabis withdrawal syndrome (CWS) in humans is characterized by various somatic symptoms, including sleep disturbances. In the present study, we investigated sleep alterations in mice after the cessation of arachidonylcyclopropylamide (ACPA), a cannabinoid type 1 receptor agonist, administration. ACPA-administered mice (ACPA mice) displayed an increased number of rearings after the cessation of ACPA administration compared to saline-administered mice (Saline mice). Moreover, the number of rubbings was also decreased in ACPA mice compared with those of the control mice. Electroencephalography (EEG) and electromyography (EMG) were measured for 3 days after the cessation of ACPA administration. During ACPA administration, there was no difference in the relative amounts of total sleep and wake time between ACPA and Saline mice. However, ACPA-induced withdrawal decreased total sleep time during the light period in ACPA mice after ACPA cessation. These results suggest that ACPA cessation induces sleep disturbances in the mouse model of CWS.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/npr2.12329

Takashi Asano

Papers

Molecular epidemiology and antimicrobial susceptibilities of 273 exfoliative toxin-encoding-gene-positive Staphylococcus aureus isolates from patients with impetigo in Japan.

Clonal change of methicillin-resistant Staphylococcus aureus isolated from patients with impetigo in Kagawa, Japan.

Cannabinoid Type 1 Receptors in the Basolateral Amygdala Regulate ACPA-Induced Place Preference and Anxiolytic-Like Behaviors

Cannabinoid Type 1 Receptors in the Basolateral Amygdala Regulate ACPA-Induced Place Preference and Anxiolytic-Like Behaviors

Sleep disturbance after cessation of cannabis administration in mice.